Human natural killer cells mediate adaptive immunity to viral antigens

Nikzad, Rana; Angelo, Laura S.; Aviles-Padilla, Kevin; Le, Duy T.; Singh, Vipul K.; Bimler, Lynn; Vukmanovic‐Stejic, Milica; Vendrame, Elena; Ranganath, Thanmayi; Simpson, Laura J.; Haigwood, Nancy L.; Blish, Catherine A.; Akbar, Arne N.; Paust, Silke

doi:10.1126/sciimmunol.aat8116

Cited by 136 publications

(145 citation statements)

References 62 publications

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“…These responses are traditionally thought to be nonspecific, as NK cell function is primarily mediated by the expression of germ-line encoded receptors rather than antigen-specific receptors [2]. However, recent evidence has revealed that NK cells are capable of antigen-specific adaptive responses against viruses, such as cytomegalovirus, Epstein-Barr Virus, Varicella Zoster Virus, and influenza virus [1, [3][4][5][6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

TIGIT is upregulated by HIV-1 infection and marks a highly functional adaptive and mature subset of natural killer cells

Vendrame

Seiler

Ranganath

et al. 2019

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Objective: Our objective was to investigate the mechanisms that govern natural killer (NK) cell responses to HIV, with a focus on specific receptor-ligand interactions involved in HIV recognition by NK cells. Design and Methods: We first performed a mass cytometry-based screen of NK cell receptor expression patterns in healthy controls and HIV + individuals. We then focused mechanistic studies on the expression and function of T cell immunoreceptor with Ig and ITIM domains (TIGIT). Results: The mass cytometry screen revealed that TIGIT is upregulated on NK cells of untreated HIV + women, but not in antiretroviral-treated women. TIGIT is an inhibitory receptor that is thought to mark exhausted NK cells; however, blocking TIGIT did not improve anti-HIV NK cell responses. In fact, the TIGIT ligands CD112 and CD155 were not upregulated on CD4 + T cells in vitro or in vivo, providing an explanation for the lack of benefit from TIGIT blockade. TIGIT expression marked a unique subset of NK cells that express significantly higher levels of NK cell activating receptors (DNAM-1, NTB-A, 2B4, CD2) and exhibit a mature/adaptive phenotype (CD57 hi , NKG2C hi , LILRB1 hi , FcRγ lo , Syk lo ). Furthermore, TIGIT + NK cells had increased responses to mock-infected and HIV-infected autologous CD4 + T cells, and to PMA/ionomycin, cytokine stimulation and the K562 cancer cell line. Conclusions: TIGIT expression is increased on NK cells from untreated HIV + individuals. Although TIGIT does not participate directly in NK cell recognition of HIV, it marks a population of mature/adaptive NK cells with increased functional responses. Antibody conjugation, mass cytometry staining and data acquisitionAntibodies were conjugated using MaxPar® ×8 labeling kits (Fluidigm). To ensure antibody stability over time, antibody panels were lyophilized into single-use pellets prior to use (Biolyph). Cells were stained for mass cytometry as described previously [51,52] and resuspended in 1× EQ Beads (Fluidigm) before acquisition on a Helios mass cytometer (Fluidigm). In vitro HIV infectionReplication competent (Q23-FL) HIV-1 was made by transfection and titrated as described [51]. CD4 + T cells were activated for 2 days with plate-bound anti-CD3 (clone OKT3, eBioscience, 10 µg/ml), soluble anti-CD28/CD49d (clones L293/L25, BD Biosciences, 1 µg/ml each) and phytohemagglutinin (eBioscience, 2.5 µg/ml). CD4 + T cells were infected overnight with Q23-FL via ViroMag R/L magnetofection (Oz Biosciences) at a multiplicity of infection of 20. HIV infection was as measured by intracellular p24. TIGIT blockade assayCells were cultured in RPMI media containing 10% FBS (Thermo Fisher Scientific) and 1% penicillin/streptomycin/amphotericin (Thermo Fisher Scientific) (RP10). NK cells were incubated in 24-well plates at a concentration of 2x10 6 cells/ml in for 3 days at 37°C with 5% CO2, with addition of rhIL-2 (R&D Systems, 300 IU/ml). After 3 days, NK cells were washed with fresh media and plated in 96-well plates (80,000-100,000 cells/well). Blocking mIgG1κ anti-h...

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Section: Introductionmentioning

confidence: 99%

TIGIT is upregulated by HIV-1 infection and marks a highly functional adaptive and mature subset of natural killer cells

Vendrame

Seiler

Ranganath

et al. 2019

Preprint

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“…While we did not find evidence of improved memory responses of peripheral blood NK cells in HIV+ individuals upon an in vitro re-stimulation with HIV-infected cells, this does not preclude the existence of these memory NK cells in other tissue locations, or in low frequency in the blood. In murine, humanized mice, and non-human primate studies of memory NK cells, memory NK cells were predominantly tissue-resident, particularly in the liver (Paust et al, 2010;Reeves et al, 2015;Nikzad et al, 2019). A low frequency of these hepatic phenotype NK cells can be found in peripheral blood (Nikzad et al, 2019) and would have been included in our analyses, but may have been at too low a frequency to detect within the bulk of the response to HIV.…”

Section: Discussionmentioning

confidence: 99%

“…In murine, humanized mice, and non-human primate studies of memory NK cells, memory NK cells were predominantly tissue-resident, particularly in the liver (Paust et al, 2010;Reeves et al, 2015;Nikzad et al, 2019). A low frequency of these hepatic phenotype NK cells can be found in peripheral blood (Nikzad et al, 2019) and would have been included in our analyses, but may have been at too low a frequency to detect within the bulk of the response to HIV. Indeed, Reeves et al have previously found that, in non-human primates, memory responses to Gag vaccination by peripheral blood NK cells were much lower in magnitude than splenic or hepatic NK cells, but were still observable (Reeves et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

“…Contrary to their classic designation as an innate immune cell type, recent work has demonstrated the ability of human NK cells to form memory against viruses including cytomegalovirus, Epstein-Barr virus and varicella-zoster virus (Gumá et al, 2004(Gumá et al, , 2006Lopez-Verges et al, 2011;Foley et al, 2012;Paust et al, 2017;Hammer et al, 2018;Nikzad et al, 2019). In non-human primates, infection with simian immunodeficiency virus (SIV) or SHIV generates antigen-specific NK cells that react with presented Gag and Env.…”

Section: Introductionmentioning

confidence: 99%

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Treated HIV Infection Alters Phenotype But Not HIV-specific Function of Peripheral Blood Natural Killer Cells

Zhao

Ferreira

Grant

et al. 2020

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ABSTRACTNatural killer (NK) cells are the predominant antiviral cells of the innate immune system, and may play an important role in acquisition and disease progression of HIV. While untreated HIV infection is associated with distinct alterations in the peripheral blood NK cell repertoire, less is known about how NK phenotype is altered in the setting of long-term viral suppression with antiretroviral therapy (ART), as well as how NK memory can impact functional responses. As such, we sought to identify changes in NK cell phenotype and function using high-dimensional mass cytometry to simultaneously analyze both surface and functional marker expression of peripheral blood NK cells in a cohort of ART-suppressed, HIV+ patients and HIV-healthy controls. We found that the NK cell repertoire following IL-2 treatment was altered in individuals with treated HIV infection compared to healthy controls, with increased expression of markers including NKG2C and CD2, and decreased expression of CD244 and NKp30. Using co-culture assays with autologous, in vitro HIV-infected CD4 T cells, we identified a subset of NK cells with enhanced responsiveness to HIV-1-infected cells, but no differences in the magnitude of anti-HIV NK cell responses between the HIV+ and HIV-groups. In addition, by profiling of NK cell receptors on responding cells, we found similar phenotypes of HIV-responsive NK cell subsets in both groups. Lastly, we identified clusters of NK cells that are altered in individuals with treated HIV infection compared to healthy controls, but found that these clusters are distinct from those that respond to HIV in vitro. As such, we conclude that while chronic, treated HIV infection induces a reshaping of the IL-2-stimulated peripheral blood NK cell repertoire, it does so in a way that does not make the repertoire more HIV-specific.

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“…Several types of memory NK cells have been characterized in humans. These include memory NK cells induced by cytokines (Cooper et al, 2009), varicella zoster virus exposure (Nikzad et al, 2019), antibodymediated stimulation (Zhang et al, 2013), or CMV-derived peptides (Hammer et al, 2018). High frequencies of NK cells expressing the activating receptor NKG2C are frequently observed in CMV seropositive individuals (Guma et al, 2006a).…”

Section: Introductionmentioning

confidence: 99%

Dynamic changes in natural killer cell subset frequencies in the absence of cytomegalovirus infection

Gyurova

Schlums

Sucharew

et al. 2019

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Individuals lacking functional natural killer (NK) cells suffer severe, recurrent infections with cytomegalovirus (CMV), highlighting the critical role of NK cells in antiviral defense. Therefore, ongoing attempts to develop an efficacious vaccine to prevent CMV infection should potentially aim to elicit NK-cell antiviral responses as an accessory to conventional T-and B-cell based approaches. In this regard, CMV infection provokes marked phenotypic and functional differentiation of the NK-cell compartment, including development of adaptive NK cells that exhibit enhanced antiviral activity. We examined longitudinal blood samples collected from 40 CMV-seronegative adolescents to ascertain whether a CMV glycoprotein B (gB) vaccine in the absence of CMV infection can stimulate differentiation or expansion of CMV-associated subsets of NK cells. Study participants uniformly lacked the CMV-dependent NKG2C + subset of NK cells, suggesting that an adjuvanted CMV gB vaccine alone is an inadequate stimulus for sustained expansion of these cells. In contrast, we observed unexpected dynamic fluctuations in the frequency of NK cells lacking FcR, EAT-2, and SYK, which were independent of vaccination or CMV infection. Whereas FcR neg NK cells in CMV infection are reported to express increased levels of the maturation marker CD57, the FcR neg NK cells observed in our CMV-negative vaccine cohort express less CD57 than their FcR + counterparts. The FcR neg NK cells in CMV-negative individuals were also functionally distinct from this subset in CMV infection, exhibiting comparable IFN- production and degranulation as FcR + NK cells in response to cytokine or antibody-dependent stimuli. These results suggest that frequencies of some NK cell subsets may increase in response to unknown environmental or inflammatory cues distinct from that which occurs after CMV infection. Greater understanding of the nature of the signals driving CMVindependent accumulation of these subsets should permit development of mechanisms to facilitate vaccine-driven expansion of CMV-reactive NK cells.Vaccine | HCMV | Innate lymphoid cell | NK cell | Memory | CD56 | CD57 | FcR | SYK | NKG2C | EAT-2

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Human natural killer cells mediate adaptive immunity to viral antigens

Cited by 136 publications

References 62 publications

TIGIT is upregulated by HIV-1 infection and marks a highly functional adaptive and mature subset of natural killer cells

TIGIT is upregulated by HIV-1 infection and marks a highly functional adaptive and mature subset of natural killer cells

Treated HIV Infection Alters Phenotype But Not HIV-specific Function of Peripheral Blood Natural Killer Cells

Dynamic changes in natural killer cell subset frequencies in the absence of cytomegalovirus infection

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